AllTaq Master Mix and PCR Core Kits
For ultrafast and versatile hot-start PCR in all applications
The AllTaq Master Mix Kit and the AllTaq PCR Core Kit provide a convenient format for highly sensitive and specific hot-start PCR using any DNA or cDNA template. Both kits are highly suited for all PCR applications and provide the following features: visual pipetting controls, gel loading and tracking dyes, an ultrafast cycling protocol, extreme room temperature stability during and after reaction setup and a 4x-concentrated master mix format, allowing for a higher sample input volume.
AllTaq Master Mix and PCR Core Kits are intended for molecular biology applications. These products are not intended for the diagnosis, prevention, or treatment of a disease.
The innovative formulation of the AllTaq Master Mix and AllTaq PCR Buffer facilitates the amplification of specific PCR products and delivers successful results at the first attempt, using the same protocol for all targets. During the annealing step of every PCR cycle, the buffer allows a high ratio of specific-to-nonspecific primer binding (see figure PCR buffer). The verified buffer composition is adapted to ultra-fast cycling conditions and simultaneously provides stringent primer-annealing conditions over a wide range of annealing temperatures. It also ensures perfect duplex capabilities. Optimization of PCR by varying the annealing temperature or the Mg2+ concentration is not required.
The versatile chemistry enables amplification of long targets up to 9 kb as well as duplex PCR, providing flexible workflow planning using the same PCR kit. Q-solution provided in the AllTaq PCR Core Kit facilitates amplification of difficult secondary structures, such as GC-rich templates. The guard-protected, hot-start mechanism prevents premature PCR leakage, ensuring premium specificity and sensitivity down to one single target molecule (see figure: Principle of AllTaq hot-start mechanism).
The AllTaq Master Mix Kit provides a convenient, ready-to-use master mix formulation, while the AllTaq PCR Core Kit offers AllTaq DNA Polymerase, AllTaq PCR buffer, dNTPs, MgCl2 and Q-solution in separate tubes, in case optimization of the PCR protocol is desired (see figure Amplification of long PCR fragments). Both kits provide orange Master Mix Tracer and blue Template Tracer in separate vials for optional use.
At low temperatures, AllTaq DNA Polymerase is kept in an inactive state by an antibody and a novel guard additive, which stabilizes the complex (see figure Principle of AllTaq hot-start mechanism). This improves the stringency of the hot start and prevents any enzymatic activity at ambient temperature, allowing reaction setup without ice or on instruments. The enzyme is fully activated after the 2-minute incubation step at 95°C and starts amplifying with high specificity from the first cycle. The hot-start procedure eliminates extension from nonspecifically annealed primers and primer–dimers in the first cycle, ensuring highly specific and reproducible PCR.
The innovative AllTaq PCR Master Mix and AllTaq Buffer facilitate ultra-fast cycling conditions and simultaneously provide stringent primer-annealing conditions over a wide range of annealing temperatures. They also ensure perfect duplex capabilities. Optimization of PCR by varying the annealing temperature or the Mg2+ concentration is not required.
The blue and orange dyes in the PCR Template Tracer and in the PCR Master Mix Tracer, respectively, allow visual tracking of pipetted samples during the PCR setup to prevent errors. When the blue template is added to the orange PCR Master Mix, the color changes to green, confirming that sample was added (see figure Visual pipetting control). The use of these tracers is optional.
Reactions can be directly loaded onto agarose gels after cycling. Each tracer dye allows monitoring of the loading process and efficient tracking of the subsequent electrophoresis.
The AllTaq Master Mix Kit provides a convenient, ready-to-use 4x concentrated master mix formulation, which allows a high template input volume to further boost sensitivity. The AllTaq PCR Core Kit offers AllTaq DNA Polymerase, PCR buffer, dNTPs, MgCl2 and Q-solution in separate tubes, if optimization of the PCR protocol should be desired.
The orange Master Mix Tracer is optionally added to master mix or PCR buffer. When template dyed with the blue Template Tracer is added to the master mix, the color of the solution changes from orange to green, providing a visual indication of correct pipetting and reaction setup (see figure PCR procedure). The tracer dyes allow direct loading on agarose gels after PCR and efficient tracking of the subsequent electrophoresis. The dyes run at approximately 50 bp (orange) and 4000 bp (blue) on a 1% agarose gel.
The versatile buffer system allows ultrafast PCR in approximately 45 minutes using the same protocol for any kind of different standard PCR, including duplex PCR. For long templates between 1–9 kb, separate cycling conditions are provided to enable amplification of long fragments using the same kit.
The guard-protected, hot-start mechanism enables complete reaction setup at room temperature, eliminating the need for ice and allowing automated pipetting. After reaction setup, samples can be left at room temperature for more than 3 days before being subjected to PCR cycling. Furthermore, the master mix is extremely stable, avoiding degradation during use and allowing storage at 2–8°C for up to 6 months.
The AllTaq PCR Core Kit also contains Q-Solution that can be added optionally. It facilitates amplification of difficult targets by modifying the melting behavior of nucleic acids (e.g., templates that have a high degree of secondary structure or that are GC rich).
To further streamline PCR workflows, we recommend the QIAgility instrument for automated reaction setup and PCR analysis on the QIAxcel Advanced system.
The AllTaq Master Mix and AllTaq PCR Core Kits can be used for any PCR for genotyping and genetic analysis or target detection from complex gDNA or cDNA templates, including reactions with multiple primer pairs or that include a wide range of amplicon sizes.
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